“Nuclease-free” Gene Editing for Sickle Cell Disease and β-Thalassemia
Faculty Information
Name:
Prof. Arun Srivastava
Email
aruns@ufl.edu
Phone
(352) 273-8259
Faculty Department/Division
Pediatrics
This project is primarily:
Basic
Research Project Description:
Human hemoglobinopathies (sickle cell disease and -thalassemia), are by far the most common (1 in 600) monogenic diseases worldwide. Although gene editing with several nucleases (mega TAL, zinc-finger nuclease, CRISPR/Cas9, base editors, prime editors) have been performed, and remarkable efficacy has been achieved in animal models, the use of these nucleases in humans has not been optimal. For example, serious adverse events, such as immune responses to these bacteria-derived nucleases, off-target cutting of DNA, chromosomal translocations, and loss of chromosomes, and genotoxicity have been observed. The lack of specificity of vehicles, such as lipid nanoparticles (LNPs), to deliver these nucleases in vivo also remains a challenge. Thus, there is need for the development alternative strategies for gene editing that are safe and efficient, as well as the development of a highly efficient and specific delivery vehicle. We have developed strategies to overcome both of these limitations. For example, we have developed a decoy zinc-finger transcription factor synthetic peptide capable of gene editing without causing double-stranded breaks in DNA. We have also identified adeno-associated virus 6 (AAV6) for high-efficiency transduction of primary human hematopoietic stem cells (HSCs). We have generated next-generation AAV6-human parvovirus B19 (B19V) hybrid and AAV-B19V chimeric vectors for high-efficiency transduction and erythroid lineage-restricted expression from B19V promoter of synthetic decoy zinc finger transcription factor peptides for “nuclease-free” gene editing for human sickle cell disease and -thalassemia. Medical students will be involved in evaluating the efficacy of further optimized AAV6-B19V vectors to achieve safe and efficacious gene editing in a xenograft mouse model. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.
Song, M.A. Kauss, E. Kopin, M. Chandra, T. Ul-Hasan, E. Miller, G.R. Jayandharan, A.E. Rivers, G.V. Aslanidi, C. Ling, B. Li, W. Ma, X. Li, L.M. Andino, L. Zhong, A.F. Tarantal, M.C. Yoder, K.K. Wong, Jr., M. Tan, S. Chatterjee, and A. Srivastava. Optimizing the transduction efficiency of capsid-modified AAV6 vectors in primary human hematopoietic stem cells in vitro and in a xenograft mouse model in vivo. Cytotherapy, 15: 986-998, 2013.
Song, X. Li, G.R. Jayandharan, Y. Wang, G.V. Aslanidi, C. Ling, L. Zhong, G. Gao, M.C. Yoder, C. Ling, M. Tan, and A. Srivastava. High-efficiency transduction of primary human hematopoietic stem cells and erythroid lineage-restricted expression by optimized AAV6 serotype vectors in vitro and in a murine xenograft model in vivo. PLoS One, 8(3): e58757, 2013.
Ling, K. Bhukhai, Z. Yin, M.Q. Tan, M.C. Yoder, P. Leboulch, E. Payen, and A. Srivastava. High-efficiency transduction of primary human CD34+ hematopoietic stem/progenitor cells by AAV6 serotype vectors: Strategies for overcoming donor-variation and implications in genome editing. Scientific Reports, 6, 35495, 2016.
Yang, K. Qing, G.D. Keeler, L. Yin, M. Mietzsch, C. Ling, B.E. Hoffman, M. Agbandje-McKenna, M. Tan, W. Wang, and A. Srivastava. Enhanced transduction of human hematopoietic stem cells by AAV6 vectors: Implications in gene therapy and genome editing. Molecular Therapy-Nucleic Acids. 20: 451-458, 2020.
Does this project have an international component or travel?
No
Neurological Complications in Fontan Patients Undergoing Cardiac Transplantation
Name: Wei Wang, MD
Email: wwang29@peds.ufl.edu
Division: Pediatric Cardiac Critical Care Medicine, Cardiology, Neurology, Pediatrics
Background:
Patients with Fontan circulation are at risk for neurological complications due to a prothrombic state, chronic microemboli, and multiple previous palliations. We describe neurological complications in patients with Fontan circulation in the early perioperative period after cardiac transplantation.
Methods:
This will be an extension of a previous retrospective, single-center case control study which evaluated the post- heart transplant neurological outcomes in all patients with Fontan physiology (4-53 years) who underwent cardiac transplantation at University of Florida. Three points of evaluation were used: (1) neurological findings on CT and MRI in the first month following transplant, (2) abnormalities on electroencephalography (EEG), and (3) the initiation of antiepileptic medications prior to post-transplant discharge. Data collected will include the patients’ preoperative anticoagulation, inotropic support, end organ function, and hemodynamics.
Role of the medical student:
Drafting and submitting a new IRB for the extension of the previous study
Making a redcap data base
Reviewing patients charts and inputting pertinent information into the RedCap database
Opportunities for poster and abstract presentations available
additional opportunities for shadowing available within the pediatric cardiac critical care unit and congenital heart center operating rooms
Med student should ideally have a strong interest in pediatric or adult cardiology or neurology. Student will work closely with Dr. Tavenner Dibert, Dr. Wei Wang and Dr. Joseph Philip from the division of pediatric cardiac critical care medicine. Students should be proactive, timely in email/text communication, and have strong communication skills. Schedule is flexible and a portion of this work may be able to be completed remotely. However, in person meetings may be required for EPIC training, patient chart review, redcap database set up etc.
Does this project have an international component or travel?
No
Next-Gen Histopathologic Evaluation of the Exocrine Pancreas in Type 1 Diabetes
Name:
Dr. Brittany Bruggeman
Email
bruggemanbr@ufl.edu
Phone
(352) 265-7337
Faculty Department/Division
Pediatrics
This project is primarily:
Translational
Research Project Description:
Type 1 diabetes is an autoimmune-mediated beta-cell disease, however, the exocrine pancreas, which controls digestion, is also significantly impacted. This project would be doing multiplexed immunofluorescence of donor pancreases from individuals with type 1 diabetes and controls to better understand what protein expression from the exocrine pancreas is most impacted, and whether this is related to insulin deficiency or exocrine pancreatic inflammation.
Students could assist with the actual procedure of staining and imaging, or assist more with the image analysis aspects once we have the images in-hand. A previous student developed a machine learning-based algorithm for image analysis, which we would work to adapt to this project. Students would be expected to be in town for most of the project, but may be able to complete part of the project remotely if we are able to set up remote imaging analysis. I would expect a publication to eventually result from this project and the student would be a named author if they contribute significantly.
Does this project have an international component or travel?
No
Low Fidelity Simulation Based Mask Ventilation Skill Drill
Name:
Dr. Wei Wang
Email
wwang29@peds.ufl.edu
Phone
(407) 431-5921
Faculty Department/Division
Pediatrics
This project is primarily:
Clinical
Research Project Description:
Simulation has become an integral part of procedural teaching for medical trainees; however, there is limited evidence in nursing education. The goal of this project is to assess the feasibility of a short low-fidelity simulation (LFS) session in improving nursing comfort in performing bag mask ventilation in critically ill children. This project will involve surveying pediatric critical care (pICU) and cardiac critical care (pCICU) nurses at Shands Children’s hospital before and after they participate in a single 5-minute low-fidelity simulation session during their clinical shifts. The sessions will be led by a PALS certified provider with scripted coaching on a pediatric airway management case scenario.
Students will be responsible for distributing pre-participation and post-participation surveys to assess the participants’ current level of experience, and comfort on performing bag-mask ventilation (BMV) based on a Likert-type scale. Students will also be responsible for inputing and analyzing survey data and writing up a subsequent manuscript for publication. Abstract presentation opportunities will be available at national level conferences such as the Society of Critical Care Medicine, as well as institutional opportunities such as the UF Patient Safety and Quality Week, Pediatrics Science Day, and UF COM Celebration of Research.
Ideally students should have an interest in medical education, simulation, pediatrics, pediatric critical care medicine, anesthesia and/or pediatric anesthesia, or emergency medicine as this project focuses on teaching of procedural techniques.
Interested students are welcome to shadow with us in the cardiac critical care unit or the cardiac operating rooms at the Congenital Heart Center.
This project cannot be done remotely as it requires in person attendance for simulation events, inputting paper survey data into Redcap, and participation in the teaching of procedures. This project has already been started and ongoing so will be easy to get involved.
Does this project have an international component or travel?
No
Prevalence of Exocrine Pancreatic Insufficiency in Adults with Type 1 Diabetes
Name:
Dr. Brittany Bruggeman
Email
bruggemanbr@ufl.edu
Phone
(352) 265-7337
Faculty Department/Division
Pediatrics
This project is primarily:
Clinical
Research Project Description:
We are currently conducting a study evaluating the prevalence of exocrine pancreatic insufficiency in adults with type 1 diabetes, and whether a survey screening tool can accurately identify these adults. The study is already IRB-approved. The student would be recruiting research participants in clinic and helping to collect data. This project would require the student to be in town for at least several days per week on average, with some remote work possible as well (data collection from Epic). We do plan to publish this study, and the student would be a listed author as long as they dedicate significant time/effort towards the project.
Does this project have an international component or travel?
No
Evaluating the prevalence and risk factors for retinopathy in pediatric type 2 diabetes
Name:
Brittany Bruggeman
Email
bruggemanbr@ufl.edu
Phone
(352) 265-7337
Faculty Department/Division
Pediatrics
This project is primarily:
Clinical
Research Project Description:
We previously collected cross-sectional data on the prevalence of retinopathy in our pediatric type 2 diabetes patient population. However, this was with a limited sample size. In order to publish results, we would like to complete a retrospective chart review of retinopathy prevalence and risk factors in our pediatric type 2 diabetes population. The student would need to complete an IRB, add on to an existing REDCap database, and collect the chart review data. This project could be completed remotely. The student would be free to be first author of the project/publication once completed.
Does this project have an international component or travel?
No
Implementation of Social Determinants of Health Screening in Pediatric Diabetes Clinic
Name:
Dr. Brittany Bruggeman
Email
bruggemanbr@ufl.edu
Phone
(321) 537-8832
Faculty Department/Division
Pediatrics
This project is primarily:
Clinical
Research Project Description:
We have previously determined that social determinants of health are prevalent in pediatric diabetes clinic. We would now like to work to implement screening utilizing Epic tools and create a referral process to either social work or our medical-legal partnership as well implement hand-out resources for social needs (previously created but need to be better implemented). This would likely be a quality improvement project that we would either publish as QI or as an advocacy project. The student would be on any future publications if they dedicated significant time/effort towards the project. Some of the project could likely be done remotely.
Does this project have an international component or travel?
No
Florida Diabetes Camp – A Model To Train the Next Generation
Name:
Dr. Paul Hiers
Email
phier88@ufl.edu
Phone
(561) 308-3639
Faculty Department/Division
Pediatrics
This project is primarily:
Case Review
Research Project Description:
This is a descriptive, retrospective study examining the distribution of demographics among the population of campers and counselors who attend and benefit from Florida Diabetes Camps (campers, counselors, staff) over the last 10 years for which there is computerized data available. We want to investigate and describe the demographic composition of campers and volunteers that support this unique camp program to see if increased diversity is observed over time. Additionally, we want to see if repeated camp attendance encourages campers to become future counselors.
Medical students role includes data input and coding, also leading to write up and future publication. Additional diabetes camp projects available as well as other diabetes projects and can discuss. Expectation to attend camp sessions during the summer as well. See potential camps and dates at FloridaDiabetesCamp.org
Does this project have an international component or travel?
No